ChIP analysis of CDP/cut binding to myeloid promoters in NB4 cells.

(A) Chromatin immunoprecipitations were performed from uninduced (0) and ATRA-induced (48 h) NB4 cells using an antibody specific for CDP/cut (lanes 1-2) and a no-antibody control (−) (lanes 3-4). The precipitated chromatin was analyzed using primers specific for the CDP/cut sites in the LF promoter, the HNP promoter, the C/EBPε promoter, and the gp91phox promoter. Input LF/CDP chromatin (1:10 dilution) is represented in lane 5. This experiment was repeated 3 times. The identities of all PCR products obtained were confirmed by dideoxy sequencing. (B) Western blot analysis was performed on 40-μg nuclear extracts prepared from uninduced (NB4; day 0), and 48-hour ATRA-induced (NB4; 48 h ATRA) NB4 cells. The blot was probed with a CDP/cut-specific antibody. The molecular-weight marker is indicated.